Heterocyclic non-nucleoside compounds, their preparation, pharmaceutical composition and their use as antiviral agents

ABSTRACT

The invention relates a kind of antiviral agents, more concretely, relates to a kind of heterocyclic non-nucleoside compounds with the following structures, their preparation and pharmaceutical compositions including the compounds. The said compounds can be used as antiviral agents and as medicaments for treating diseases such as hepatitis B, influenza, herpes, HIV and so on.

CROSS-REFERENCED TO RELATED APPLICATIONS

This is a national stage of International Application No.PCT/CN2007/001861, filed on Jun. 13, 2007, which claims priority ofChinese Patent Application No. 200610027724.0, filed Jun. 13, 2006, thedisclosure of each application is hereby incorporated by reference intheir entirety.

TECHNICAL FIELD

The present invention relates to a kind of antiviral inhibitors,particularly to a kind of heterocyclic non-nucleoside compounds, theirpreparing methods, and pharmaceutical compositions including the same.The said compounds can be used as antiviral inhibitors and asmedicaments for treating diseases such as hepatitis B, influenza,herpes, AIDS and so on.

BACKGROUND ART

Human pathogenic viruses are a kind of nucleic acid particles with verysimple structure. Most of them lack enzymatic system, and have to dependon the host cells to replicate their nucleic acids and proteins and thenassemble into virus particles so as to realize the multiplication of theviruses. Viral infection can cause various diseases and harm the healthand lives of human severely. With the undercount, about 60% epidemicaldiseases are caused by viral infection. Up to now, more than 3000 kindsof viruses have been found all over the world, and new viruses are beingcontinuously found. At the World Virology Convention held in Paris inAugust 2002, the 7^(th) report of International Committee on theTaxonomy of Viruses embodies more than 3600 kinds of viruses, in whichmore than 1200 kinds of viruses are pathogenic to human and can bedivided into 29 families, 7 subfamilies and 53 genera. At present, theviruses with high morbidity and great harmfulness mainly includeinfluenza virus, hepatitis B virus, AIDS virus, cytomegalovirus andherpes virus, etc.

Now there still lacks of drugs with high specificity in the treatment ofviral diseases, and also the commonly used drugs in clinical therapymainly are divided into the following types: the antiviral drugs forinhibiting viral replication; the immunomodulators for enhancing thebody's immune function; the antitussive, anodyne, antipyretic andantiphlogistic and the like against clinical symptoms; theanti-infection drugs for preventing secondary infection; the vaccinesfor preventing viral infection and the disinfectants for blocking thetransmission of viruses, etc.

The study of new drugs for treating viral diseases abroad focuses on theantiviral drugs. At present, the anti-influenza virus drugs include theadamantine amine drugs, the neuraminidase inhibitors of influenza virus,the receptor blocking agent of influenza virus and the antisenseoligonucleotide against influenza virus etc. And the ones used inclinical therapy mainly are the adamantanamine drugs and theneuraminidase inhibitors. However, the hepatitis virus infection is awell-known difficulty in therapeutics so far. More than 80% of the acuteinfection of the hepatitis B virus (HBV), hepatitis C virus (HCV) andhepatitis D virus (HDV) will convert into chronic infection, in which20% persistent infection may develop to hepatic cirrhosis, and 1% to 5%will change to hepatoma carcinoma. The world health organization hasclassified hepatitis as the 9th cause of death. China is a region withhigh incidence of viral hepatitis, and the carriers of hepatitis B virusare more than 120 million. It is estimated that the direct economic lossresulted from viral hepatitis is 30 billion to 50 billion RMB.Therefore, it is a main task for the medical and pharmaceutical domesticand abroad scientists to explore and develop antiviral drugs. Thevidarabine, vidarabine phosphate, acyclovir, zidovudine studied in1980's are not used to treat hepatitis B now abroad due to their poortherapeutic effect and strong toxic adverse effect. In recent years,many large-scale enterprises developed various nucleoside drugs havingobvious inhibitory effect to HBV, such as lamivudine, famciclovir,lobucavir, adefovir dipivoxiil, FTC (dideoxyfluorinethiocytosine), FMAU(fluoro-methylarabinosyluracil), FDDC (fluoro-dideocytosine), BMS 200475(epoxyhydroxylcarbodeoxy guanosine), by using the established hepatomacarcinoma cell lines, hepatitis virus transfected cell lines ortransgenic cell lines, and transgenic mouse hepatitis animal model toscreen the drugs against hepatitis B virus and hepatitis C virus. Theresearchers abroad carried out the preclinical research on more than 30kinds of drugs in 1998-2002. And recently there are 21 drugs enteringstage II-III clinical trial, and among them, the trial medicaments foranti-hepatitis B virus are mostly derived from the HIV revertaseinhibitor and herpes virus DNA polymerase inhibitor, in which enticavirhas been authorized to be commercially available in 2005. Most of thetrail medicaments for anti-hepatitis C virus are derived frombroad-spectrum antiviral drugs or RNA virus inhibitors and theimmunomodulators having antiviral activity.

At present, most of the authorized antivirus drugs are nucleosidecompounds. During the clinical uses, they have the followingdisadvantages: 1) cytotoxicity; 2) occurrence of drug resistant virusesvariants induced by long-term medication and the requirement of theother drugs having different structure to antagonize these variants.Therefore, developing non-nucleoside antiviral drugs becomes anarresting aspect.

DISCLOSURE OF THE INVENTION

An object of the present invention is to design and synthesize a kind ofnew heterocyclic non-nucleoside compounds as antiviral inhibitors so asto pave the way for finding precursor compounds or antiviral drugs forthe research of antiviral drugs.

Another object of the present invention is to provide methods forpreparing heterocyclic non-nucleoside compounds of the presentinvention.

Still another object of the present invention is to provide apharmaceutical composition containing the heterocyclic non-nucleosidecompounds of the present invention as active components.

Further another object of the present invention is to provide uses ofthe heterocyclic non-nucleoside compounds of the present invention inpreparing medicaments for treating viral diseases.

The heterocyclic non-nucleoside compounds provided by the presentinvention have a structure represented by the following formula:

wherein, X₁ is NR₄, O or S;

R₁ is pyridyl, substituted pyridyl, phenyl, substituted phenyl,5-membered heterocyclic group, substituted 5-membered heterocyclic groupor fused heterocyclic group, wherein the 5-membered heterocyclic groupis one containing one or two hetero atoms selected from the groupconsisting of N, O and S, and the fused heterocyclic group is quinolylor indolyl;

Each R₂ and R₃ is one independently selected from the group consistingof H; halogen atom; nitro group; C₁-C₂₅ alkyl; C₁-C₂₅ alkyl substitutedby halogen atom, C₁-C₂₅ alkyl, C₃-C₂₅ cycloalkyl, C₁-C₂₅ alkoxyl, C₃-C₂₅cycloalkoxyl, C₂-C₂₅ alkenyl, C₃-C₂₅ cycloalkenyl, nitro group, aminogroup, C₁-C₂₅ alkylamino group, C₃-C₂₅ cycloalkylamino group, C₁-C₂₅alkyl amide group, hydroxyl group, acyloxy or C₁-C₄ alkoxycarbonyl;hydroxyl group; amino group; C₁-C₂₅ alkylamino group; C₃-C₂₅cycloalkylamino group; C₃-C₉ cycloalkyl; C₃-C₉ cycloalkyl substituted byhalogen atom, C₁-C₂₅ alkyl, C₃-C₂₅ cycloalkyl; C₁-C₂₅ alkoxyl, C₃-C₂₅cycloalkoxyl, C₂-C₂₅ alkenyl, C₃-C₂₅ cycloalkenyl, nitro group, aminogroup, C₁-C₂₅ alkylamino group, C₃-C₂₅ cycloalkylamino group, C₁-C₂₅alkyl amide group, hydroxyl or acyloxy; aryl; aryl substituted byhalogen atom, C₁-C₂₅ alkyl, C₃-C₂₅ cycloalkyl, C₁-C₂₅ alkoxyl, C₃-C₂₅cycloalkoxyl, C₂-C₂₅ alkenyl, C₃-C₂₅ cycloalkenyl, nitro group, aminogroup, C₁-C₂₅ alkylamino group, C₃-C₂₅ cycloalkylamino group, C₁-C₂₅alkyl amide group, hydroxyl or acyloxy; benzyl; benzyl substituted byhalogen atom, C₁-C₂₅ alkyl, C₃-C₂₅ cycloalkyl, C₁-C₂₅ alkoxyl, C₃-C₂₅cycloalkoxyl, C₂-C₂₅ alkenyl, C₃-C₂₅ cycloalkenyl, nitro group, aminogroup, C₁-C₂₅ alkylamino group, C₃-C₂₅ cycloalkylamino group, C₁-C₂₅alkyl amide group, hydroxyl or acyloxy; C₂-C₂₅ alkenyl; C₂-C₂₅ alkenylsubstituted by halogen atom, C₁-C₂₅ alkyl, C₃-C₂₅ cycloalkyl, C₁-C₂₅alkoxyl, C₃-C₂₅ cycloalkoxyl, nitro group, amino group, C₁-C₂₅alkylamino group, C₃-C₂₅ cycloalkylamino group, C₁-C₂₅ alkyl amidegroup, hydroxyl, acyloxy or C₁-C₄ alkoxycarbonyl; C₃-C₂₅ cycloalkenyl;C₃-C₂₅ cycloalkenyl substituted by halogen atom, C₁-C₂₅ alkyl, C₃-C₂₅cycloalkyl, C₁-C₂₅ alkoxyl, C₃-C₂₅ cycloalkoxyl, nitro group, aminogroup, C₁-C₂₅ alkylamino group, C₃-C₂₅ cycloalkylamino group, C₁-C₂₅alkyl amide group, hydroxyl or acyloxy;

wherein each R′ and R″ independently is hydrogen atom, C₁-C₂₅ alkyl,halogenated C₁-C₁₀ alkyl, aryl, benzyl, substituted benzyl, C₁-C₆hydroxyalkyl, substituted or unsubstituted heterocyclic methylene,wherein the substituents on benzyl may be halogen, C₁-C₁₀ alkyl, C₁-C₁₀alkoxyl, C₁-C₁₀ alkylamino group, nitrile group, carboxyl or C₁-C₁₀alkoxycarbonyl; and ureido group or ureylene;

R₄ is H, aryl, substituted aryl, benzyl, C₁-C₁₃ alkyl, substitutedC₁-C₁₃ alkyl, C₂-C₆ alkenyl or C₃-C₆ cycloalkyl, wherein thesubstituents on the said aryl or C₁-C₁₃ alkyl may be halogen atom,alkoxyl, amino group, alkylamino group or hydroxyl.

In one preferable embodiment of the present invention, the said X₁ isNR₄, R₄ is hydrogen atom or C₁-C₄ alkyl;

R₁ preferably is pyridyl, substituted pyridyl, thiazolyl, substitutedthiazolyl, quinolyl or indolyl, wherein the substituent(s) of thesubstituted pyridyl and substituted thiazolyl is one or two selectedfrom the group of hydroxyl, halogen atom, nitro group, C₁-C₄ alkyl,C₁-C₄ alkoxyl, phenyl, benzyl and C₁-C₄ alkoxycarbonylamino group; andR₁ further preferably is 2-pyridyl, 2-thiazolyl, 2-quinolyl or2-indolyl;

R₂ preferably is linear or branched C₁-C₈ alkyl or C₃-C₈ cycloalkyl,more preferably linear or branched C₁-C₆ alkyl or C₃-C₆ cycloalkyl, suchas methyl, ethyl, propyl, butyl, pentyl, hexyl, cyclopropyl, cyclobutyl,cyclopentyl or cyclohexyl, most preferably isobutyl or cyclohexyl;

R₃ is

wherein each R′ and R″ independently is hydrogen atom, C₁-C₂₅ alkyl,halogenated C₁-C₁₀ alkyl, aryl, benzyl, halogenated benzyl, benzylsubstituted by C₁-C₁₀ alkyl, benzyl substituted by C₁-C₁₀ alkoxyl,benzyl substituted by C₁-C₁₀ alkylamino group, benzyl substituted bynitrile group, benzyl substituted by carboxyl, benzyl substituted byC₁-C₁₀ alkoxycarbonyl, substituted or unsubstitutedheterocyclomethylene; more preferably R₃ is

wherein R′ is C₁-C₆ linear or branched alkyl, and preferably C₁-C₄linear or branched alkyl, most preferably ethyl;

In this embodiment, the representative specific compound of the presentinvention is one of the following compounds:

In another preferable embodiment of the present invention,

X₁ is O;

R₁ preferably is pyridyl, substituted pyridyl, quinolyl or indolyl,wherein the said substituted pyridyl is pyridyl substituted by one ortwo substituents selected from hydroxyl, halogen atom, nitro group,C₁-C₄ alkyl, C₁-C₄ alkoxyl, phenyl, benzyl and C₁-C₄ alkoxycarbonylaminogroup; and R₁ further preferably is 2-pyridyl, 2-quinolyl or 2-indolyl;

R₂ preferably is linear or branched C₁-C₈ alkyl or C₃-C₈ cycloalkyl,more preferably linear or branched C₁-C₆ alkyl or C₃-C₆ cycloalkyl, suchas methyl, ethyl, propyl, butyl, pentyl, hexyl, cyclopropyl, cyclobutyl,cyclopentyl or cyclohexyl, most preferably isobutyl;

R₃ is

wherein each R′ and R″ independently is hydrogen atom, C₁-C₂₅ alkyl,halogenated C₁-C₁₀ alkyl, aryl, benzyl, halogenated benzyl, benzylsubstituted by C₁-C₁₀ alkyl, benzyl substituted by C₁-C₁₀ alkoxyl,benzyl substituted by C₁-C₁₀ alkylamino group, benzyl substituted bynitrile group, benzyl substituted by carboxyl, benzyl substituted byC₁-C₁₀ alkoxycarbonyl, substituted or unsubstitutedheterocyclomethylene; more preferably R₃ is

wherein R′ is linear or branched C₁-C₆ alkyl, and preferably linear orbranched C₁-C₄ alkyl, most preferably ethyl;

In this embodiment, the representative specific compound of the presentinvention is one of the following compounds:

In still another preferable embodiment of the present invention,

X₁ is S;

R₁ preferably is pyridyl, substituted pyridyl, phenyl, substitutedphenyl, thiazolyl, substituted thiazolyl, quinolyl or indolyl; whereinthe said substituted pyridyl, substituted phenyl and substitutedthiazolyl are respectively pyridyl, phenyl and thiazolyl each of whichare substituted by one or two substituents selected from hydroxy,halogen atom, nitro group, C₁-C₄ alkyl, C₁-C₄ alkoxyl, phenyl, benzyland C₁-C₄ alkoxycarbonylamino group; and R₁ further preferably ispyridyl; pyridyl substituted by hydroxy; phenyl; phenyl substituted byone or two substituents selected from hydroxyl, halogen atom, nitrogroup, C₁-C₄ alkyl, C₁-C₄ alkoxyl, phenyl and C₁-C₄ alkoxycarbonylaminogroup; 2-thiazolyl; 2-thiazolyl substituted by halogen atom, phenyl orC₁-C₄ alkyl; 2-quinolyl or 2-indolyl;

Most preferably, the said heterocyclic non-nucleoside compounds have astructure represented by the following formula I:

Wherein each R₅ and R₆ independently is hydrogen atom, halogen atom,linear or branched C₁-C₄ alkyl or phenyl;

R₂ preferably is linear or branched C₁-C₈ alkyl, C₃-C₈ cycloalkyl orbenzyl, more preferably linear or branched C₁-C₆ alkyl, C₃-C₆ cycloalkylor benzyl, such as methyl, ethyl, propyl, butyl, pentyl, hexyl,cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl, most preferablyisobutyl, n-butyl or benzyl;

R₃ is one selected from hydrogen atom; C₁-C₆ hydroxyalkyl; C₁-C₄ alkylsubstituted by C₁-C₄ alkoxycarbonyl; C₂-C₄ alkenyl substituted by C₁-C₄alkoxycarbonyl;

wherein each R′ and R″ independently is hydrogen atom, C₁-C₂₅ alkyl,halogenated C₁-C₁₀ alkyl, aryl, benzyl, halogenated benzyl, benzylsubstituted by C₁-C₁₀ alkyl, benzyl substituted by C₁-C₁₀ alkoxy, benzylsubstituted by C₁-C₁₀ alkylamino group, benzyl substituted by nitrilegroup, benzyl substituted by carboxyl, benzyl substituted by C₁-C₁₀alkoxycarbonyl, substituted or unsubstituted heterocyclomethylene.

Further preferably, R₃ is one group selected from hydrogen atom; C₁-C₄hydroxyalkyl; ethoxycarbonylethylene; ethoxycarbonylvinyl;

wherein R′ is linear or branched C₁-C₄ alkyl;

wherein R′ is hydrogen atom or linear or branched C₁-C₄ alkyl;

wherein each R′ and R″ independently is hydrogen atom, linear orbranched C₁-C₄ alkyl, phenyl, benzyl, fluorobenzyl or4-[2-(2-thiazolyl)-5-isobutyl-thiazolyl]methylene;

In this embodiment, the representative specific compound of the presentinvention is one of the following compounds:

The heterocyclic non-nucleoside compounds of the present invention maybe prepared by the following method:

The compounds (compounds 4i) with X₁ being NR₄ (R₄ being H) of thepresent invention can be prepared according to the following chemicalreaction equation:

Wherein:

R₁ is pyridyl, substituted pyridyl, phenyl, substituted phenyl,5-membered heterocyclic group, substituted 5-membered heterocyclicgroup, quinolyl or indolyl, wherein the 5-membered heterocyclic group isa 5-membered heterocyclic group containing one or two hetero atomsselected form N, O and S;

R₂ is one selected from the group consisting of C₁-C₂₅ alkyl; C₁-C₂₅alkyl substituted by halogen atom, C₁-C₂₅ alkyl, C₃-C₂₅ cycloalkyl,C₁-C₂₅ alkoxyl, C₃-C₂₅ cycloalkoxyl, C₂-C₂₅ alkenyl, C₃-C₂₅cycloalkenyl, nitro group, amino group, C₁-C₂₅ alkylamino group, C₃-C₂₅cycloalkylamino group, C₁-C₂₅ alkyl amide group, hydroxyl, acyloxy orC₁-C₄ alkoxycarbonyl; C₃-C₉ cycloalkyl; C₃-C₉ cycloalkyl substituted byhalogen atom, C₁-C₂₅ alkyl, C₃-C₂₅ cycloalkyl, C₁-C₂₅ alkoxyl, C₃-C₂₅cycloalkoxyl, C₂-C₂₅ alkenyl, C₃-C₂₅ cycloalkenyl, nitro group, aminogroup, C₁-C₂₅ alkylamino group, C₃-C₂₅ cycloalkylamino group, C₁-C₂₅alkyl amide group, hydroxyl or acyloxy; aryl; aryl substituted byhalogen atom, C₁-C₂₅ alkyl, C₃-C₂₅ cycloalkyl, C₁-C₂₅ alkoxyl, C₃-C₂₅cycloalkoxyl, C₂-C₂₅ alkenyl, C₃-C₂₅ cycloalkenyl, nitro group, aminogroup, C₁-C₂₅ alkylamino group, C₃-C₂₅ cycloalkylamino group, C₁-C₂₅alkyl amide group, hydroxyl or acyloxy; benzyl; benzyl substituted byhalogen atom, C₁-C₂₅ alkyl, C₃-C₂₅ cycloalkyl, C₁-C₂₅ alkoxyl, C₃-C₂₅cycloalkoxyl, C₂-C₂₅ alkenyl, C₃-C₂₅ cycloalkenyl, nitro group, aminogroup, C₁-C₂₅ alkylamino group, C₃-C₂₅ cycloalkylamino group, C₁-C₂₅alkyl amide group, hydroxy or acyloxy; C₂-C₂₅ alkenyl; C₂-C₂₅ alkenylsubstituted by halogen atom, C₁-C₂₅ alkyl, C₃-C₂₅ cycloalkyl, C₁-C₂₅alkoxyl, C₃-C₂₅ cycloalkoxyl, nitro group, amino group, C₁-C₂₅alkylamino group, C₃-C₂₅ cycloalkylamino group, C₁-C₂₅ alkyl amidegroup, hydroxyl, acyloxy or C₁-C₄ alkoxycarbonyl; C₃-C₂₅ cycloalkenyl;C₃-C₂₅ cycloalkenyl substituted by halogen atom, C₁-C₂₅ alkyl, C₃-C₂₅cycloalkyl, C₁-C₂₅ alkoxyl, C₃-C₂₅ cycloalkoxyl, nitro group, aminogroup, C₁-C₂₅ alkylamino group, C₃-C₂₅ cycloalkylamino group, C₁-C₂₅alkyl amide group, hydroxyl or acyloxy;

R₃ is H, C₁-C₂₅ alkyl or

wherein R′ is C₁-C₂₅ alkyl and the like;

Wherein, the compound 1 is commercially available, for example, fromSinopharm Chemical Reagent Co. Ltd. and Aldrich Co. etc. The compound 2can be synthesized according to prior art of J. Org. Chem. 1973; 38;3571.

In the presence of N-ethyl-N′-(3-dimethylaminopropyl) carbodiimidechloride (EDC), N,N-dimethylpyridine (DMAP) and molecular sieve, in thesolvent of DMA, DMF, acetonitrile, dichloromethane or tetrahydrofuran,and under the alkaline condition of pyridine, N-methyl morpholine,triethylamine or diethylpropylethylamine etc., the reaction of thecompound 1 and the compound 2 is carried out to produce intermediatecompound 3.

Compound 3 is heated to 120-140° C. in the mixture of ammonium acetateand sodium acetate and produces compound 4i through ring closingreaction.

The compounds (compounds 4ii) with X₁ being S of the present inventioncan be prepared according to the following chemical reaction equation:

Wherein the Lawesson's reagent is2,4-bis(para-methoxyphenyl)-1,3-dithio-2,4-bisulfide, which iscommercially available. The Lawesson's reagent is added to the aboveobtained compound 3 in the solvent of tetrahydrofuran, then heated andrefluxed to produce the compound 4ii.

The compounds (compounds 4iii) with X₁ being O of the present inventioncan be prepared according to the following chemical reaction equation:

The phosphorus oxychloride solution including the above obtainedcompound 3 is heated to 100-130□ and reacted to produce the compound4iii.

The compound 4 with R₃ being CO₂Et can react according to the followingchemical reaction equation to obtain the compound 5:

The compound 4 can be the compound 4i, 4ii or 4iii.

Under alkaline condition of lithium hydroxide, sodium hydroxide etc. orunder acidic condition of hydrochloric acid, sulfuric acid etc., thecompound 4 is hydrolyzed to obtain the compound 5. The condition of thereaction may be room temperature or heated to 100□.

The compound 6 can be prepared from the compound 5 according to thefollowing chemical reaction equation:

Wherein each R′ and R″ independently is hydrogen atom, C₁-C₂₅ alkyl,halogenated C₁-C₁₀ alkyl, aryl, benzyl, substituted benzyl, C₁-C₆hydroxyalkyl, substituted or unsubstituted heterocyclomethylene, whereinthe substitutent on benzyl group may be halogen, C₁-C₁₀ alkyl, C₁-C₁₀alkoxyl, C₁-C₁₀ alkylamino group, nitrile group, carboxyl or C₁-C₁₀alkyloxycarbonyl; X is O or NH.

The compound 5 is transformed into acyl chloride under the condition ofoxalyl chloride or thionyl chloride etc, and then acyl chloride isreacted with various alcohols, various substituted amine or aqueousammonia etc. to produce the compound 6. The solvent of the reaction maybe dichloromethane, ethyl acetate, water or the mixture thereof. Thereaction is conducted under alkaline condition, and the alkaline reagentmay be inorganic alkali such as potassium carbonate, potassiumbicarbonate, sodium carbonate or sodium bicarbonate etc., or organicalkali such as pyridine, N-methylmorpholine, isobutyl chloroformate,triethylamine and diethylpropylethylamine etc.

Or the compound 7 can be prepared from the compound 4 according to thefollowing chemical reaction equation:

Wherein, the compound 4 reacted to produce compound 7 by adding aluminumlithium hydride in the organic solvent of anhydrous tetrahydrofuran orabsolute ether etc. under the temperature of −20□-25□.

Or the compound 8 can be prepared from the compound 7 according to thefollowing chemical reaction equation:

Wherein, IBX is 2-iodoxybenzoic acid, which is purchased from AldrichCo. The Grignard reagent is C₁-C₂₅ alkyl or C₃-C₈ cycloalkyl Grignardreagent, i.e., R is C₁-C₂₅ alkyl or C₃-C₈ cycloalkyl. The compound 7 isoxidized to produce intermediate compound 8. The compound 8 reacted toproduce the compound 9 by adding Grignard reagent in solvent of absoluteether or anhydrous tetrahydrofuran. The compound 4 can also conduct thisreaction instead of the compound 8.

Or the compound 10 can be prepared from the compound 9 according to thefollowing chemical reaction equation:

The compound 9 can be oxidized to produce the compound 10 by using theactive MnO₂ in acetone solution or IBX in the mixed solution of tolueneand DMSO etc.

Or the compound 19 can be prepared according to the following chemicalreaction equation:

Wherein, R₁, R₂, R₃ are defined as same as above, X is halogen. Thesynthesis of the compound 13 can refer to the prior art (J. Chem. Soc.Perkin I. 1982, 159-164; Heterocyclic. Chem. 1991, 28, 1003). Thecompound 12 reacted for 0.5-2 hours by adding butyllithium in organicsolvent of anhydrous tetrahydrofuran or absolute ether, after that,anhydrous zinc chloride is added to produce zincon. Or, active zincpowder is added into the organic solvent of anhydrous tetrahydrofuran orabsolute ether including compound 12 and the above mixture is heated andrefluxed to produce zincon. And then the compound 13 is added and reactfor 8-24 hours to obtain the compound 19 in the solvent of benzene ortoluene and the like, under the presence of the palladium catalyst(tetra(triphenylphosphine)palladium, or the mixture of palladium acetateand triphenyl phosphine).

Or the compound 14 and the compound 15 can be prepared according to thefollowing chemical reaction equation:

Wherein, R is C₁-C₂₅ alkyl, phenyl or

wherein R′ is C₁-C₂₅ alkyl etc. The compound 20 is commerciallyavailable, e.g. from Sinopharm Chemical Reagent Co. Ltd, Aldrich Co.etc. The compound 8 and the compound 20 react to produce the compound 14in anhydrous tetrahydrofuran solution by adding NaH, and the compound 14is hydrogenated to obtain the compound 15.

Or the compound 16 can be prepared according to the following chemicalreaction equation:

Wherein, the compound 17 is the compound 4 with R₁ being 2-thiazolyl;R₂, R₃ are defined as same as above; R is C₁-C₂₅ alkyl or C₃-C₈cycloalkyl. The compound 17 reacted for 24 hours to produce compound 18in dichloromethane solution by adding liquid bromine. The compound 18reacts under room temperature to produce compound 16 in the solvent ofDMA or DMF by adding the zincon and catalyst nickel.

Or the compound 19 can be prepared according to the following chemicalreaction equation:

Wherein, R₁, R₂, R₃ are defined as same as above. The boron reagent canbe commercially available, e.g. from Sinopharm Chemical Reagent Co. Ltd,Aldrich Co. etc. The synthesis method of the compound 13 can refer tothe prior art (J. Chem. Soc. Perkin I. 1982, 159-164; Heterocyclic.Chem. 1991, 28, 1003). The boron reagent and the compound 13 react for12 hours to obtain compound 19 in the solvent of methanol or ethanol byadding palladium catalyst, water and alkali of potassium carbonate orsodium carbonate.

Or the compound 21 can be prepared according to the following chemicalreaction equation:

Wherein, R₄ is aryl, substituted aryl, benzyl, C₁-C₁₃ alkyl, substitutedC₁-C₁₃ alkyl, C₂-C₆ alkenyl or C₃-C₆ cycloalkyl, wherein thesubstituent(s) on the said aryl or C₁-C₁₃ alkyl may be halogen atom,alkoxyl, amino group, alkylamino group or hydroxyl; X is halogen atom.

The compound 4i reacts for 1 hour at room temperature by adding NaH inanhydrous tetrahydrofuran solution, and then halide is added and reactsfor 12 hours to obtain compound 21.

Generally TLC is used to detect the completion degree of the reaction.After the reaction completed, the reaction is typically quenched withice-water, and the resultant mixture is extracted with ethylether, ethylacetate, dichloromethane, trichloromethane etc., and washed with 5%hydrochloric acid, water, saturated brine in sequence, then dried, andthe solvent is removed under reduced pressure at low temperature. Thefinal product is obtained through column chromatography and isidentified by the methods of nuclear magnetic resonance (NMR), massspectrum and the like.

The present invention provides an antiviral pharmaceutical composition,which contains one or more of the above heterocyclic non-nucleosidecompounds as active compounds, and may further contains pharmaceuticallyconventional adjuvant, such as excipient, disintegrant, anti-oxidant,sweetening agent, coating agent etc.

Advantageous Effect

The present invention designs and synthesizes a kind of new heterocyclicnon-nucleoside antiviral agents, which can effectively inhibit thereplication of influenza virus, the DNA replication of hepatitis B virus(HBV), and the formation of HBsAg and HBeAg. These compounds can be usedfor the preparation of medicaments for treating viral diseases, and mayovercome the disadvantages of the known nucleosides drugs, includingcytotoxicity, and the requirement of other drugs having differentstructure against the drug-resistant virus variants induced by long-termmedication. The structure of the compounds according to the invention isrelatively simple and easy to be prepared.

DESCRIPTION OF THE DRAWING

FIG. 1 is the comparison of the inhibition ratios of DHBV-DNA level induck serum of the treating group and that of the control group withviral infection, after the duck hepatitis B virus infected ducks wereorally (intragastric) administrated.

BEST MODE OF THE INVENTION

The present invention will be further described in conjugation with thefollowing specific examples, but the present invention is not limitedthereto.

PREPARATION EXAMPLES

In the following preparation examples, NMR was measured using Mercury-Vx300M produced by Varian, and the NMR calibration was: δH/C 7.26/77.0 ppm(CDCl₃). The reagents were provided mainly by Shanghai Chemical ReagentCo., Ltd. The purification of the products were performed mainly withcolumn chromatography of silica gel (200-300 mesh). And the type of thesilica gel used in column chromatography was coarse hollow (ZLX-□),which was produced by the Branch of Qingdao Haiyang Chemical Plant.

Preparation Example 1

The compound 1.1 (3.3 mmol), the compound 2.1 (3 mmol),N-ethyl-N′-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) (3.3mmol), N,N-dimethylpyridine (DMAP) (0.3 mmol) and molecular sieve weremixed, and cooled in ice bath (0□). Then DMF (5 mL), pyridine (4.5 mmol)were added sequentially. The completion degree of the reaction wastracked by TLC. After the reaction was completed, the reaction solutionwas diluted with water (25 mL), extracted with EtOAc (25 mL). Thesolvent was removed completely by concentration. Then the compound 3.1was obtained by separation through column chromatography with petroleumether/ethyl acetate (volume ratio 5:1). Then the compound 3.1 (0.6 mmol)was mixed with ammonium acetate (NH₄OAc) (15 mmol), sodium acetate(NaOAc) (30 mmol) and heated to 130□. The completion degree of thereaction was tracked by TLC. Then the reaction solution was cooled toroom temperature and diluted with water (50 mL), extracted with ethylacetate (50 mL). The solvent was removed completely by concentration.Then the compound 4i.1 was obtained by separation through columnchromatography with petroleum ether/ethyl acetate (volume ratio 1:1).

Except pyridine-2-carboxylic acid was replaced by the substitutedcarboxylic acid (compound 1) listed in the following table and thecompound 2.1 was replaced by various compound 2, the following compoundswere synthesized by the same method as preparation example 1:

Compound Compound Compound Structure formula ¹H NMR (CDCl₃, 4i 1 2 ofcompound 4i 300 MHz) data C273

δ 0.97 (d, J = 6.6 Hz, 6H), 1.42 (t, J = 7.2 Hz, 3H), 2.06 (m, 1H), 2.88(d, J = 7.2 Hz, 2H), 4.39 (q, J = 7.2 Hz, 2H), 7.31 (t, J = 9.0 Hz, 1H),7.83 (t, J = 7.8 Hz, 1H), 8.28 (d, J = 7.8 Hz, 1H), 8.53 (d, J = 4.2 Hz,1H). C311-2

δ 1.04 (d, J = 6.6 Hz, 6H), 1.44 (t, J = 7.2 Hz, 3H), 2.02 (m, 1H), 2.94(d, J = 7.2 Hz, 2H), 4.34 (q, J = 6.9 Hz, 2H), 7.11 (s, 1H), 7.21 (t, J= 7.2 Hz, 1H), 7.26 (t, J = 7.2 Hz, 1H), 7.48 (d, J = 3.3 Hz, 1H), 7.56(d, J = 7.8 Hz, 1H), 10.90 (br, 1H). C323-2

δ 0.94 (d, J = 6.6 Hz, 6H), 1.36 (t, J = 7.2 Hz, 3H), 1.92 (m, 1H), 2.75(d, J = 7.2 Hz, 2H), 4.35 (q, J = 6.9 Hz, 2H), 7.50 (t, J = 7.8 Hz, 1H),7.64 (t, J = 6.9 Hz, 1H), 7.79 (d, J = 7.8 Hz, 1H), 7.94 (d, J = 8.4 Hz,1H), 8.23 (d, J = 8.1 Hz, 1H), 8.42 (d, J = 8.4 Hz, 1H). C305-2

δ 0.95 m, 6H), 1.70 (m, 5H), 1.96 (m, 2H), 3.20 (m, 1H), 4.37 (t, 2H),7.36 (d, 1H), 7.80 (d, 1H), 11.10 (bs, 1H).

Preparation Example 2

The compound 3.1 (0.6 mmol) and Lawesson's reagent (0.9 mmol) weremixed, then after adding THF (5 mL), heated and refluxed. The completiondegree of the reaction was tracked by TLC. Subsequently the reactionsolution was cooled to room temperature, and concentrated to remove thesolvent completely. The compound 4ii.1 was obtained by separationthrough column chromatography with petroleum ether/ethyl acetate (volumeratio 3:1).

Except pyridine-2-carboxylic acid was replaced by the substitutedcarboxylic acid (compound 1) listed in the following table and thecompound 2.1 was replaced by the different compound 2, the variouscompound 3 can be synthesized by the same method as preparationexample 1. Then the following target compounds were synthesize by thesame method as preparation example 2:

compound compound compound structure formula ¹H NMR (CDCl₃, 4ii 1 2 ofcompound 4ii 300 MHz) data C290

δ 0.92 (d, J = 6.6 Hz, 6H), 1.35 (t, J = 7.2 Hz, 3H), 1.94 (m, 1H), 3.07(d, J = 7.2 Hz, 2H), 4.36 (q, J = 7.2 Hz, 2H), 7.22 (t, J = 5.1 Hz, 1H),7.69 (t, J = 1.8 Hz, 1H), 8.17 (d, J = 7.5 Hz, 1H), 8.48 (d, J = 4.2 Hz,1H). W28

δ 0.99 (d, 6H), 1.99 (m, 1H), 3.15 (d, 2H), 3.96 (s, 3H), 7.46 (d, 1H),7.86 (d, 1H). C328-2

δ 1.04 (d, J = 6.6 Hz, 6H), 1.28 (t, J = 7.2 Hz, 3H), 2.02 (m, 1H), 3.16(d, J = 7.2 Hz, 2H), 4.34 (q, J = 6.9 Hz, 2H), 6.94 (s, 1H), 7.10 (t, J= 6.9 Hz, 1H), 7.21 (m, 2H), 7.62 (d, J = 8.1 Hz, 1H), 10.17 (br, 1H).C340

δ 1.02 (d, J = 6.6 Hz, 6H), 1.36 (t, J = 7.2 Hz, 3H), 2.06 (m, 1H), 3.18(d, J = 7.2 Hz, 2H), 4.45 (q, J = 6.9 Hz, 2H), 7.53 (t, J = 7.8 Hz, 1H),7.71 (t, J = 6.9 Hz, 1H), 7.80 (d, J = 7.8 Hz, 1H), 8.08 (d, J = 8.4 Hz,1H), 8.22 (d, J = 8.1 Hz, 1H), 8.38 (d, J = 8.4 Hz, 1H). C289

δ 0.98(d, J = 6.6 Hz, 6H), 1.40 (t, J = 7.2 Hz, 3H), 1.97 (m, 1H), 3.11(d, J = 7.2 Hz, 2H), 4.45 (q, J = 6.9 Hz, 2H), 7.38 (br, 3H), 7.92 (br,2H). C290-2

δ 0.98 (d, J = 6.6 Hz, 6H), 1.41 (t, J = 7.2 Hz, 3H), 1.98 (m, 1H), 3.13(d, J = 7.2 Hz, 2H), 4.41 (q, J = 7.2 Hz, 2H), 7.35 (dd, J1 = 5.1 Hz, J2= 4.8 Hz, 1H), 8.24 (d, J = 8.1 Hz, 1H), 8.62 (d, J = 4.8 Hz, 1H), 9.09(s, 1H). C290-3

δ 0.95 (d, J = 6.6 Hz, 6H), 1.38 (t, J = 7.2 Hz, 3H), 1.94 (m, 1H), 3.10(d, J = 7.2 Hz, 2H), 4.38 (q, J = 7.2 Hz, 2H), 7.75 (dd, J = 4.8 Hz,2H), 8.63 (d, J = 4.5 Hz, 1H). C306-3

δ 1.02 (d, J = 6.6 Hz, 6H), 1.43 (t, J = 7.2 Hz, 3H), 2.05 (m, 1H), 3.18(d, J = 7.2 Hz, 2H), 4.40 (q, J = 7.2 Hz, 2H), 7.27 (t, J = 5.1 Hz, 1H),7.49 (d, J = 3.3 Hz, 1H), 8.16 (d, J = 4.5 Hz, 1H), 11.72 (bs, 1H).C306-2

δ 0.94 (d, J = 6.6 Hz, 6H), 1.42 (t, J = 7.2 Hz, 3H), 1.93 (m, 1H), 3.08(d, J = 7.2 Hz, 2H), 4.42 (q, J = 7.2 Hz, 2H), 7.67 (m, 2H), 8.06 (d, J= 7.5 Hz, 1H). C218

δ 0.98 (d, J = 6.6 Hz, 6H), 1.93 (m, 1H), 2.74 (d, J = 7.2 Hz, 2H), 7.28(q, J = 5.7 Hz, 2H), 7.57 (s, 1H), 7.77 (t, J = 7.8 Hz, 1H), 8.14 (d, J= 8.1 Hz, 1H), 8.59 (d, J = 4.8 Hz, 1H). C375

δ 1.00 (d, J = 6.6 Hz, 6H), 1.44 (t, J = 7.2 Hz, 3H), 2.00 (m, 1H), 3.15(d, J = 7.2 Hz, 2H), 4.42 (q, J = 6.9 Hz, 2H), 7.74 (s, 1H). C307-2

δ 0.96 (d, J = 6.6 Hz, 6H), 1.39 (t, J = 7.2 Hz, 3H), 1.95 (m, 1H), 3.09(d, J = 7.2 Hz, 2H), 4.39 (q, J = 7.2 Hz, 2H), 7.07 (m, 2H), 7.88 (m,2H). C307-3

δ 0.94 (d, J = 6.6 Hz, 6H), 1.38 (t, J = 7.2 Hz, 3H), 1.94 (m, 1H), 3.08(d, J = 7.2 Hz, 2H), 4.37 (q, J = 7.2 Hz, 2H), 7.06 (m, 1H), 7.33 (m,1H), 7.63 (m, 2H). C307-4

δ 1.00 (d, J = 6.6 Hz, 6H), 1.45 (t, J = 7.2 Hz, 3H), 2.02 (m, 1H), 3.15(d, J = 7.5 Hz, 2H), 4.44 (q, J = 7.2 Hz, 2H), 7.22 (m, 2H), 7.40 (m,1H), 8.34 (m, 1H). C319-3

δ 0.97 (d, J = 6.6 Hz, 6H), 1.41 (t, J = 7.2 Hz, 3H), 1.96 (m, 1H), 3.09(d, J = 7.2 Hz, 2H), 3.82 (s, 3H), 4.40 (q, J = 7.2 Hz, 2H), 6.91 (d, J= 8.7 Hz, 2H), 7.85 (d, J = 8.4 Hz, 2H). C319-4

δ 1.00 (d, J = 6.6 Hz, 6H), 1.44 (t, J = 7.2 Hz, 3H), 1.99 (m, 1H), 3.13(d, J = 7.2 Hz, 2H), 3.87 (s, 3H), 4.43 (q, J = 7.2 Hz, 2H), 6.95 (d, J= 8.4 Hz, 1H), 7.34 (t, J = 8.4 Hz, 1H), 7.49 (d, J = 7.8 Hz, 1H), 7.52(s, 1H). C319-5

δ 0.99 (d, J = 6.6 Hz, 6H), 1.44 (t, J = 7.2 Hz, 3H), 2.03 (m, 1H), 3.14(d, J = 7.2 Hz, 2H), 3.99 (s, 3H), 4.43(q, J = 7.2 Hz, 2H), 6.98 (d, J =8.4 Hz, 1H), 7.07 (t, J = 7.5 Hz, 1H), 7.35 (t, J = 8.1 Hz, 1H), 8.45(d, J = 7.8 Hz, 1H). C334-2

δ 0.99 (d, J = 6.6 Hz, 6H), 1.42 (t, J = 7.2 Hz, 3H), 1.98 (m, 1H), 3.14(d, J = 7.2 Hz, 2H), 4.42 (q, J = 7.2 Hz, 2H), 8.08 (d, J = 6.9 Hz, 2H),8.25 (d, J = 6.9 Hz, 2H). C334-3

δ 0.96 (d, J = 6.6 Hz, 6H), 1.40 (t, J = 7.2 Hz, 3H), 1.95 (m, 1H), 3.11(d, J = 7.2 Hz, 2H), 4.38 (q, J = 7.2 Hz, 2H), 7.58 (t, J = 8.1 Hz, 1H),8.21 (t, J = 7.5 Hz, 2H), 8.68 (s, 1H). C334-4

δ 1.01 (d, J = 6.6 Hz, 6H), 1.40 (t, J = 7.2 Hz, 3H), 2.00 (m, 1H), 3.17(d, J = 7.5 Hz, 2H), 4.39 (q, J = 7.2 Hz, 2H), 7.66 (m, 3H), 7.93 (d, J= 7.8 Hz, 1H). C404

δ 0.96 (d, J = 6.6 Hz, 6H), 1.38 (t, J = 7.2 Hz, 3H), 1.45 (s, 9H), 1.95(m, 1H), 3.09 (d, J = 6.9 Hz, 2H), 4.38 (q, J = 7.2 Hz, 2H), 7.11 (s,1H), 7.44 (d, J = 8.7 Hz, 2H), 7.81 (d, J = 8.4 Hz, 2H). C323-3

δ 1.00 (d, J = 6.6 Hz, 6H), 1.44 (t, J = 7.2 Hz, 3H), 1.99 (m, 1H), 3.14(d, J = 7.2 Hz, 2H), 4.44 (q, J = 7.2 Hz, 2H), 7.37 (m , 2H), 7.77 (d, J= 6.9 Hz, 1H), 7.96 (s, 1H). C358

δ 1.02 (d, J = 6.6 Hz, 6H), 2.02 (m, 1H), 3.18 (d, J = 7.2 Hz, 2H), 3.97(s, 3H), 7.44 (m, 3H), 7.62 (d, J = 8.1 Hz, 2H), 8.03 (s, 1H). C303-1

δ 0.99 (d, J = 6.6 Hz, 6H), 1.43 (t, J = 7.2 Hz, 3H), 1.99 (m, 1H), 2.36(s, 3H), 3.12 (d, J = 7.2 Hz, 2H), 4.42 (q, J = 7.2 Hz, 2H), 7.20 (d, J= 8.1 Hz, 2H), 7.82 (d, J = 8.4 Hz, 2H). C303-2

δ 1.00 (d, J = 6.6 Hz, 6H), 1.44 (t, J = 7.2 Hz, 3H), 1.99 (m, 1H), 2.39(s, 3H), 3.13 (d, J = 7.2 Hz, 2H), 4.43 (q, J = 7.2 Hz, 2H), 7.21 (d, J= 7.5 Hz, 1H), 7.29 (t, J = 7.8 Hz, 1H), 7.70 (d, J = 7.8 Hz, 1H), 7.80(s, 1H). C303-3

δ 0.99 (d, J = 6.6 Hz, 6H), 1.40 (t, J = 7.2 Hz, 3H), 2.00 (m, 1H), 2.55(s, 3H), 3.15 (d, J = 7.2 Hz, 2H), 4.40 (q, J = 7.2 Hz, 2H), 7.24 (m,3H), 7.64 (d, J = 7.5 Hz, IH). C364-2

δ 1.01 (d, J = 6.6 Hz, 6H), 1.45 (t, J = 7.2 Hz, 3H), 2.01 (m, 1H), 3.16(d, J = 7.2 Hz, 2H), 4.08 (s, 3H), 4.45 (q, J = 7.2 Hz, 2H), 7.50 (d, J= 8.4 Hz, 1H), 7.75 (s, 1H), 7.92 (d, J = 8.4 Hz, 1H). C305-4

δ 0.99 (d, J = 6.6 Hz, 6H), 1.34 (t, J = 7.2 Hz, 3H), 1.98 (m, 1H), 2.36(s, 3H), 3.10 (d, J = 7.2 Hz, 2H), 4.35 (q, J = 7.2 Hz, 2H), 6.80 (d, J= 8.7 Hz, 2H), 7.78 (d, J = 9.0 Hz, 2H), 8.11 (s, 1H). C364-1

δ 0.96 (d, J = 6.6 Hz, 6H), 1.39 (t, J = 7.2 Hz, 3H), 1.95 (m, 1H), 3.12(d, J = 7.2 Hz, 2H), 3.90 (s, 3H), 4.35 (q, J = 7.2 Hz, 2H), 7.10 (d, J= 7.8 Hz, 1H), 7.44 (t, J = 8.1 Hz, 1H).

Preparation Example 3

The compound 3.1 (0.6 mmol) and POCl₃ (3 mL) were mixed and heated to80□. The completion degree of the reaction was tracked by TLC. Then thereaction solution was decanted to saturated NaHCO₃ solution (50 mL) of0□, and POCl₃ was removed. Then the resulting solution was extractedwith ethyl acetate (50 mL), and the solvent was removed completely byconcentration. The compound 4iii.1 was obtained by separation throughcolumn chromatography with petroleum ether/ethyl acetate (volume ratio4:1).

Except pyridine-2-carboxylic acid was replaced by the substitutedcarboxylic acid (compound 1) listed in the following table, the variouscompounds 3 can be synthesized by the same method as preparationexample 1. Then the following target compounds were synthesized by thesame method as preparation example 3:

structure Compound Compound formula of ¹H NMR (CDCl₃, 4iii 1 compound4iii 300 MHz) data C274

δ 0.92 (d, J = 6.6 Hz, 6H), 1.35 (t, J = 7.2 Hz, 3H), 2.13 (m, 1H), 2.96(d, J = 7.2 Hz, 2H), 4.36 (q, J = 7.2 Hz, 2H), 7.30 (t, J = 5.1 Hz, 1H),7.73 (t, J = 7.8 Hz, 1H), 8.17 (d, J = 7.8 Hz, 1H), 8.48 (d, J = 4.8 Hz,1H). C324-4

δ 1.01 (d, J = 6.6 Hz, 6H), 1.42 (t, J = 7.2 Hz, 3H), 2.25 (m, 1H), 3.09(d, J = 7.2 Hz, 2H), 4.43 (d, J = 6.9 Hz, 2H), 7.59 (t, J = 7.2 Hz, 1H),7.73 (t, J = 7.2 Hz, 1H), 7.77 (d, J = 8.4 Hz, 1H), 8.27 (m, 3H).

Preparation Example 4

The compound 4.1 (0.5 mmol) and lithium hydroxide (LiOH) (2 mmol) weremixed, and the mixed solvent of MeOH (4 mL) and water (1 mL) was added.The reaction was conducted at room temperature and tracked by TLC. Afterthe reaction was completed, the solvent was concentrated, and thereaction solution was acidified with 1 mol/L hydrochloric acid (10 mL),extracted with ethyl acetate (25 mL). The solvent was removed completelyby concentration to obtain the compound 5.1.

The following compound can be synthesized by the same method.

¹H NMR (CDCl₃, 300 compound structure formula MHz) data Wang268-1

δ 0.11 (d, 6H), 2.03 (m, 1H), 3.33 (d, 2H), 7.40 (d, 1H), 7.80 (d, 1H).Wang268

δ 0.76 (t, 3H), 1.13 (m, 2H), 1.38 (m, 2H), 2.47 (t, 2H), 7.36 (d, 1H),7.74 (d, 1H). Wang302

δ 4.89 (s, 2H), 7.27 (5H), 7.38 (d, 1H), 7.80 (d, 1H).

Preparation Example 5

The compound 5.1 (0.5 mmol) was dissolved in dichloromethane (25 mL),and oxalyl Chloride (0.6 mmol) was added. After the resulting mixturewas spin dried, ethyl acetate and water (10 mL:10 mL) were added, andsodium bicarbonate (1.0 mmol) was added, then para-fluorobenzylamine(0.6 mmol) was added. After the resulting mixture was agitated at roomtemperature for several hours, water (10 mL) was added, and extractedwith ethyl acetate (25 mL) twice. The organic phase was washed with 1Nhydrochloric acid (20 mL) twice and saturated saline one time, and driedover MgSO₄, and then the organic phase was concentrated. The mixture waspurified through chromatographic column with petroleum ether/ethylacetate (volume ratio 4:1) to obtain the product 6.1.

Except para-fluorobenzylamine was replaced by various substituted amineor alcohol listed in the following table, the following compounds weresynthesized by the same method as preparation example 5:

Substituted structure ¹H NMR (CDCl₃, compound amine or alcohol formula300 MHz) data W28F

δ 1.02 (d, J = 6.6 Hz, 6H), 2.03 (m, 1H), 3.31 (d, J = 7.2 Hz, 2H), 4.61(d, J = 6.0 Hz, 2H), 7.04 (m, 1H), 7.45 (t, J = 8.1 Hz, 2H), 7.35 (m,2H), 7.44 (d, J = 3.0 Hz, 1H), 7.82 (m, 1H), 7.88 (d, J = 3.0 Hz, 1H).C267 NH₃

δ 1.01 (d, J = 6.6 Hz, 6H), 2.02 (m, 1H), 3.28 (d, J = 6.9 Hz, 2H), 5.60(bs, 2H), 7.43 (d, J = 3.0 Hz, 1H), 7.88 (d, J = 3.0 Hz, 1H). C357

δ 1.02 (d, J = 6.6 Hz, 6H), 2.04 (m, 1H), 3.31 (d, J = 7.2 Hz, 2H), 4.64(d, J = 6.3 Hz, 2H), 7.34 (bm, 5H), 7.85 (d, J = 3.3 Hz, 2H). C343

δ 1.02 (d, J = 6.6 Hz, 6H), 2.04 (m, 1H), 3.31 (d, J = 7.2 Hz, 2H), 4.64(d, J = 6.3 Hz, 2H), 7.34 (bm, 4H), 7.85 (d, J = 3.3 Hz, 1H). C295-3

δ 0.93 (d, J = 6.6 Hz, 6H), 1.90 (m, 1H), 2.84 (d, J = 6.9 Hz, 2H), 3.02(s, 3H), 3.09 (s, 3H), 7.39 (d, J = 3.3 Hz, 1H), 7.82 (d, J = 3.0 Hz,1H). C281

δ 0.98 (d, J = 6.6 Hz, 6H), 1.99 (m, 1H), 2.98 (d, J = 4.8 Hz, 3H), 3.26(d, J = 7.2 Hz, 2H), 7.42 (d, J = 3.3 Hz, 1H), 7.44 (bs, 1h), 7.85 (d, J= 3.3 Hz, 1H). C310-4 (W28D)

δ 1.00 (d, 6H), 1.42 (d, 6H), 2.00 (m, 1H), 3.12 (d, 3H), 5.29 (m, 1H),7.45 (d, 1H), 7.86 (d, 1H). C324-6

δ 0.93( d, J = 6.6 Hz, 6H), 1.64 (s, 9H), 1.93 (m, 1H), 3.04 (d, J = 7.5Hz, 2H), 7.40 (d, J = 3.0 Hz, 1H), 7.80 (d, J = 3.0 Hz, 1H). C503

δ 1.01 (m, 12H), 1.93 (m, 1H), 2.05 (m, 1H), 2.81 (d, J = 6.9 Hz, 2H),3.31 (d, J = 7.2 Hz, 2H), 4.70 (d, J = 4.2 Hz, 2H), 7.04 (m, 1H), 7.44(d, J = 3.0 Hz, 2H), 7.88 (d, J = 3.0 Hz, 2H), 8.18 (bs, 1H). W28MF

δ 1.01 (d, J = 6.3 Hz, 6H), 2.01 (m, 1H), 2.52 (s, 3H), 3.29 (d, J = 6.9Hz, 2H), 4.60 (d, J = 5.4 Hz, 2H), 7.05 (d, J = 8.1 Hz, 2H), 7.34 (m,2H), 7.52 (s, 1H), 7.78 (bs, 1H).

Preparation Example 6

The compound 4.1 (0.5 mmol) was dissolved in anhydrous THF (25 mL), andaluminum lithium hydride (LiAlH₄) (0.6 mmol) was added. The reaction wasconducted in ice bath and tracked by TLC. After the reaction wascompleted, the reaction solution was diluted with water (25 mL), andextracted with ethyl acetate (25 mL). The solvent was removed completelyby concentration. The compound 7.1 was obtained by separation throughchromatographic column with petroleum ether/ethyl acetate (volume ratio4:1).

¹H NMR (CDCl₃, 300 compound structure formula MHz) data C254

δ O.96 (d, J = 6.6 Hz, 6H), 1.88 (m, 1H), 2.71 (d, J = 7.2 Hz, 2H), 2.91(bs, 1H), 4.70 (s, 2H), 7.38 (d, J = 3.0 Hz, 1H), 7.83 (d, J = 3.3 Hz,1H).

Preparation Example 7

The compound 7.1 (1 mmol) and IBX (1.25 mmol) were mixed, and the mixedsolvent of toluene and DMSO (2 mL:1 mL) was added. The reaction wasconducted at 50□ and tracked by TLC. After the reaction was completed,the reaction solution was extracted with ethyl acetate (25 mL), and thesolvent was removed completely to obtain the compound 8.1. The compound8.1 was dissolved in absolute ether (10 mL), and methyl Grignard reagent(1.2 mmol) was added dropwise. The reaction was conducted at roomtemperature and tracked by TLC. After the reaction was completed, thereaction solution was diluted with water (25 mL), and extracted withethyl acetate (25 mL), then the solvent was removed completely byconcentration. The compound 9.1 was obtained by separation throughchromatographic column with petroleum ether/ethyl acetate (volume ratio2:1).

Except methyl Grignard reagent was replaced by ethyl Grignard reagent,the following compound C282-2 was synthesized by the same method aspreparation example 7:

¹H NMR (CDCl₃, 300 compound structure formula MHz) data C268-2

δ 0.96 (d, J = 6.6 Hz, 6H), 1.57 (d, J = 6.6 Hz, 3H), 1.90 (m, 1H), 2.70(d, J = 7.2 Hz, 2H), 3.07 (bs, 1H), 4.95 (m, 1H), 7.39 (d, J = 3.0 Hz,1H), 7.84 (d, J = 3.0 Hz, 1H), H). C282-2

δ 0.96 (m, 9H), 1.90 (d, J = 6.6 Hz, 3H), 2.68 (d, J = 7.2 Hz, 2H), 4.64(m, 1H), 7.38 (d, J = 3.3 Hz, 1H), 7.82 (d, J = 3.0 Hz, 1H).

Preparation Example 8

The compound 9.1 (0.5 mmol) and IBX (0.6 mmol) were mixed, and the mixedsolvent of toluene and DMSO (2 mL:1 mL) was added. The reaction wasconducted at 50□ and tracked by TLC. After the reaction was completed,the reaction solution was diluted with water (25 mL), and extracted withethyl acetate (25 mL), and then the solvent was removed completely byconcentration to obtain the compound 10.1. The following compound C280-4can be synthesized from compound C282-2 by the same method:

¹H NMR (CDCl₃, 300 compound structure formula MHz) data C266

δ 0.99 (d, J = 6.6 Hz, 6H), 1.99 (m, 1H), 2.71 (s, 3H), 3.17 (d, J = 6.9Hz, 2H), 7.45 (d, J = 3.3 Hz, 1H), 7.89 (d, J = 3.0 Hz, 1H). C280-4

δ 0.98 d, J = 6.6 Hz, 6H), 1.20 (t, J = 7.2 Hz, 2H), 1.99 (m, 1H), 3.17(m, 4H), 7.44 (d, J = 3.3 Hz, 1H), 7.86 (d, J = 3.0 Hz, 1H).

Preparation Example 9

The compound 12.1 (1.0 mmol) was dissolved in anhydrous THF, and nBuLi(1.05 mmol) was added dropwise at −78□. After the reaction was conductedfor half an hour, anhydrous zinc chloride (1.1 mmol) was added. Afterthe reaction solution was agitated for half an hour at room temperature,the compound 13.1 (11.0 mmol) and catalyst (0.05 mmol) were added, andthe benzene as solvent (10 mL) was added and reacted for 12 hours at80□. The reaction was tracked by TLC. After the reaction was completed,the solvent was removed completely by concentration. The product wasdiluted with water (25 mL), and extracted with ethyl acetate (25 mL).Then the solvent was removed completely by concentration. The compound11.1 was obtained by separation through chromatographic column withpetroleum ether/ethyl acetate (volume ratio 4:1).

¹H NMR (CDCl₃, 300 compound structure formula MHz) data C276

δ 0.98 (d, J = 6.6 Hz,6H), 2.00 (m, 1H), 3.16 (d, J = 7.2 Hz, 2H), 3.94(s, 3H), 7.32 (t, J = 5.1 Hz, 1H), 779 (t, J = 1.8 Hz, 1H), 8.27 (d, J =7.5 Hz, 1H), 8.58 (d, J = 4.2 Hz, 1H).

Preparation Example 10

The compound 20.1 (1.05 mmol) was dissolved in anhydrous THF (10 mL),and NaH (1.05 mmol) was added at 0□. After the reaction was conductedfor half an hour, the compound 8.1 (1.0 mmol) was added and reacted for12 hours at 0□ under stirring. The reaction was tracked by TLC. Afterthe reaction was completed, the solvent was removed completely byconcentration. The product was diluted with water (25 mL) and extractedwith ethyl acetate (25 mL). Then the solvent was removed completely byconcentration. The compound 14.1 was obtained by separation throughchromatographic column with petroleum ether/ethyl acetate (volume ratio4:1). The compound 14.1 was dissolved in MeOH (10 mL) with adding 10%Pd/C and hydrogenated at room temperature overnight. The compound 15.1was obtained by vacuum filtration.

¹H NMR (CDCl₃, 300 compound structure formula MHz) data C322-2

δ 0.99 (d, J = 6.6 Hz, 6H), 1.35 (t, J = 7.2 Hz, 3H), 1.95 (m, 1H), 2.84(d, J = 7.5 Hz, 2H), 4.29 (q, J = 7.2 Hz, 2H), 6.90 (d, J = 15.3 Hz,1H), 7.44 (d, J = 3.0 Hz, 1H), 7.63 (d, J = 15.3 Hz, 1H), 7.88 (d, J =3.0 Hz, 1H). C324-5

δ 0.95 (d, J = 6.6 Hz, 6H), 1.23 (t, J = 7.2 Hz, 3H), 1.87 (m, 1H), 2.67(d, J = 7.2 Hz, 3H), 2.76 (t, J = 7.2 Hz, 2H), 2.98 (t, J = 7.2 Hz, 2H),4.13 (q, J = 7.2 Hz, 2H), 7.34 (d, J = 3.0 Hz, 1H), 7.81 (d, J = 3.0 Hz,1H).

Preparation Example 11

The compound 17.1 (1.0 mmol) was dissolved in DCM (10 mL) with addingliquid bromine (1.2 mmol). The reaction was conducted for 12 hours atroom temperature and tracked by TLC. After the reaction was completed,the solvent was removed completely by concentration. The remainder wasextracted with ethyl acetate (25 mL), and the compound 18.1 was obtainedby separation through chromatographic column with petroleum ether/ethylacetate (volume ratio 4:1). The iodomethane (1.2 mmol) and active zincpowder (1.3 mmol) were reacted in DMA to obtain zincon (1.2 mmol), thencompound 18.1 and catalyst (10%) were added. After the reaction wasconducted for 3 hours at room temperature, the reaction solution wasdiluted with water (25 mL) and extracted with ethyl acetate (25 mL).Then the solvent was removed completely by concentration to obtain thecrude product. The crude product was separated through chromatographiccolumn with petroleum ether/ethyl acetate (volume ratio 4:1) to obtainthe compound 16.1.

¹H NMR (CDCl₃, 300 compound structure formula MHz) data W28M

δ 0.97 (d, J = 6.6 Hz, 6H), 1.39 (d, J = 6.3 Hz, 6H), 1.95 (m, 1H), 2.50(s, 3H), 3.09 (d, J = 7.2 Hz, 2H), 5.26 (m, 1H), 7.50 (s, 1H).

Preparation Example 12

The para-methoxybenzeneboronic acid reagent (1.0 mmol) was dissolved inethanol (10 mL), then adding water (2 mL), potassium carbonate (3.0mmol), compound 21.1 (1.0 mmol) and the catalyst (0.05 mmol). Thereaction was conducted for 12 hours at 80□ and tracked by TLC. After thereaction was completed, the solvent was removed completely byconcentration. The remainder was diluted with water (25 mL), andextracted with ethyl acetate (25 mL). The solvent was removed completelyby concentration, and the compound 19.1 was obtained by separationthrough chromatographic column with petroleum ether/ethyl acetate(volume ratio 5:1).

¹H NMR (CDCl₃, 300 compound structure formula MHz) data C37

δ 0.99 (d, J = 5.4 Hz, 6H), 1.41 (d, J = 6.0 Hz, 6H), 1.98 (m, 1H), 3.09(d, J = 7.2 Hz, 2H), 3.85 (s, 3H), 5.28 (m, 1H), 6.93 (d, J = 8.7 Hz,2H), 7.89 (d, J = 9.0 Hz, 2H).

Preparation Example 13

The compound 4i.1 (1.0 mmol) was dissolved in anhydrous tetrahydrofuran(10 mL), and NaH (1.2 mmol) was added. After the reaction was conductedfor 1 hour at room temperature, iodomethane (3.0 mmol) was added, afterthat the reaction further was conducted for 12 hours at roomtemperature. The reaction was tracked by TLC. After the reaction wascompleted, the reaction solution was concentrated to remove the solventcompletely, and acidified with 5% hydrochloric acid, diluted with water(20 mL), then extracted with ethyl acetate (25 mL). The solvent wasremoved completely by concentration, and the compound 21.1 was obtainedby separation through chromatographic column with petroleum ether/ethylacetate (volume ratio 5:1).

¹H NMR (CDCl₃, 300 compound structure formula MHz) data C319-6

δ 1.48 (m, 5H), 1.73 (m, 4H), 1.83 (m, 4H), 3.20 (m, 1H), 4.35 (s, 3H),4.37 (q, 2H), 7.39 (d, 1H), 7.89 (d, 1H).

Experimental Example Experimental Example 1 Experiment of Anti HepatitisB Virus (HBV) Activity Test

1. Object of the Experiment:

The sample compounds are screened for their anti hepatitis B virus (HBV)activity. The experiment includes: testing the effect of cytotoxicity ofthe sample compounds on secretion of the surface antigen and coreantigen of hepatitis B virus and on replication of virus nucleic acid(DNA) through the virus-cell level experiment.

2. Principle of the Experiment:

Hepatitis B virus (HBV) transgenic human hepatoma carcinoma cell,HepG2.2.15 cell line, can secrete hepatitis B virus particles(containing antigen and DNA) into the supernatant when being incubated.

Under the influence of antiviral drugs, the contents of HBsAg, HBeAg andviral DNA secreted from cells into the supernatant were detected.Compared with the contents of control groups without drugs, theantiviral activity of the sample drugs can be observed. Meanwhile, thecytotoxic effect of the sample drugs can be detected. The valueconcentration of the sample drugs which caused 50% of the cells deadmeasured with MTT method was CC₅₀. And the value concentration of thesample drugs which inhibited 50% of the secretion of HBsAg and HBeAgdetected by specific ELISA kits and inhibited 50% of the viral DNAreplication detected by fluorescent quantitation PCR method was IC₅₀.

3. Samples of the Experiment:

The solutions of the sample drugs having required concentration (1 mM)were prepared before use. Each sample drug was tested with 7 dilutionconcentrations, and the antiviral drugs such as Lamivudine etc. wereused as the positive control drugs to check whether each experiment wasnormal or not each time.

4. Method of the Experiment:

a) Experimental Process and Collection of Supernatant

HepG2.2.15 cells were inoculated into a 96-well plate, and the sampledrugs were added next day. The medium and the sample drugs with the sameconcentration were renewed periodically, and the supernatants werecollected on the eighth day to be detected. MTT was added to the cellsin the 96-well plate, and MTT dissolving solution was added after 4hours and reacted overnight. OD₅₇₀ was measured on ELISA the next day.The cytotoxicity of the sample drugs to HepG2.2.15 cells, the effect ofthe sample drugs on the growth of the cells, and the concentration ofthe sample drugs (CC₅₀) resulting in 50% of the cells dead wereestimated according to the OD values.

b) Detection of the Contents of HBsAg and HBeAg in Supernatant (ELISAMethod):

HBsAg and HBeAg were detected with reagent kits (Purchased from Sinθ-American Biotechnological Company). The samples were added to thecoated strip plate, and the same amount of enzyme labeled conjugate wasadded. After reacted for one hour at 37□, the plate was washed 5 times.The colorant Solutions A and B were added, and the reaction was stoppedafter 15 minutes, and OD_(450/630) was measured. The half inhibitionconcentration (IC₅₀) of the sample for the HBV antigen was estimatedaccording to the OD values.

c) Detection of the Content of HBV-DNA in Supernatant by FluorescentQuantification PCR:

A suitable amount of supernatant was added to the same volume of virusextract solution, and was boiled after mixed homogenously, thencentrifuged at 10000 rpm under room temperature for 5 minutes, and asuitable amount of supernatant was collected for PCR amplification.Meanwhile five HBV-DNA standard samples were set to establish a standardcurve. According to the obtained viral DNA replication value, theinhibition ratios of HBV-DNA replication of each sample drug atdifferent concentrations were calculated, and then the half inhibitionrates of the sample drugs were calculated to obtain IC₅₀. For thosesamples that can not be calculated for IC₅₀ values, they can berepresented by ICx and list the corresponding concentration values.

The PCR primers used in the experiment were:

P1: 5′ATCCTGCTGCTATGCCTCATCTT3′ P2: 5′ ACAGTGGGGAAAGCCCTACGAA3′.

The PCR probe used in the experiment was:

5′TGGCTAGTTTACTAGTGCCATTTTG3′

5. Experimental Results:

Cyto- HbsAg HBeAg DNA toxicity secretion secretion replication SampleCC₅₀ IC₅₀ (SI) IC₅₀ (SI) IC₅₀/IC_(X) (SI) No. (μM) (μM) (μM) (μM)C290 >100 NT NT 0.14 (>714) C290-3 >100 NT NT NA C306-2 >100 NT NT 0.14(>714) C306-3 5.67 NT NT 0.14 (>40.5) C290-2 >100 NT NT NA C274 >100 NTNT 67.6 (>1.5) C323-2 >100 NT NT NA C324-4 45.4 NT NT 11.9 (4)C375 >33.3 NT NT >33.3 C307-2 >33.3 NT NT NA C307-3 >33.3 NT NT <0.05(>666) C307-4 >33.3 NT NT 0.14 (>237.9) C319-3 >33.3 NT NT 0.09 (>370)C319-4 >33.3 NT NT 0.11 (>302.7) C319-5 >33.3 NT NT 0.04 (>832.5)C334-2 >33.3 NT NT 1.06 (>31.4) C334-3 >33.3 NT NT 1.2 (>27.75)C334-4 >33.3 NT NT 1.24 (>26.9) C404 >33.3 NT NT NA C323-3 >33.3 NT NTNA C254 >33.3 NT NT >33.3 C358 >33.3 NT NT IC36 = 1.23 C303-1 >33.3 NTNT 2.9 (>11.5) C303-2 >33.3 NT NT >33.3 C303-3 >33.3 NT NT 1.48 (>22.5)C296-4 >33.3 NT NT 0.2 (>166.5) C364-1 19.2 NT NT 1.1 (18) C364-2 >33.3NT NT NA C305-4 >33.3 NT NT NA C268-1 >33.3 NT NT 1.3 (>25.6)C282-1 >33.3 NT NT <0.05 (>666) C268-2 >33.3 NT NT 0.6 (>55.5)C282-2 >33.3 NT NT 0.8 (>41.6) C266 >33.3 NT NT 0.06 (>555) C280-4 >33.3NT NT 0.09 (>370) C322-2 >33.3 NT NT 0.06 (>555) C324-5 >33.3 NT NT 3(>11.1) C324-6 >33.3 NT NT 3.7 (>9) C310-4 >100 NT NT 0.81 (>123) (W28D)W28F 43 NT NT 0.78 (55) W28M >20 NT NT <0.08 (>250) W28 >333 67.57(>4.93) 21.67 (>15.4) 2.4 (>138.8) C37 >33 NT NT 0.03 (>1100) Wang268256.7 37.2 (6.9)  127.7 (2)    IC53 = 125  Wang302 192.2 NC 83.2 (2.3) 63.1 (3.1) C295-3 >100 NT NT IC12 = 1.23 C281 >100 NT NT IC94 = 33  C343 50 NT NT IC19 = 1.23 C267 >100 NT NT IC89 = 33   Note: CC₅₀indicates the effect of the sample drugs on the growth of HepG2.2.15cells, 50% death concentration. IC₅₀ is the concentration of sampledrugs that inhibits 50% of the antigen or DNA replication. SI is theselection index of the bioactivity of the samples. SI value >2 meanseffective and the greater SI value is better. NA means no obviousbiological activity or unable to be calculated. NT means no test. NCmeans no results.

It can be seen from the experiment results that most of this kind ofcompounds have inhibition activity to HBV DNA replication on cellularlevel, wherein IC₅₀ values of 19 compounds were less than 1 μM, and IC₅₀values of 8 compounds were less than 0.1 μM.

Experimental Example 2 Experiment of Anti Hepatitis B Virus (HBV)Activity Test on Animal

1. Drugs

The solutions of the sample drugs having required concentration (100mg/mL) were prepared by using 0.3% sodium carboxymethyl cellulose. Batchnumber: 00701010

Positive drug, Lamivudine, a product of Glaxo Wellcome pharmaceuticalCo., was prepared with normal saline. Batch number: 005110011

2. Virus:

Strong positive serum of Duck hepatitis B virus DNA (DHBV-DNA),collected from Shanghai brown-duck, was store at −70□.

3. Animals:

1-day-old Beijing duck, purchased from Beijing QianJin breeding duckfarm.

4. Reagents

α-³² P-dCTP was purchased from Beijing FuRui Biotechnology Co. Nicktranslation kit was purchased from Promega Co.; Sephadex G-50, FicollPVP was purchased from Pharmacia Co. (Sweden); SDS was purchased fromMerck Co. (Germany); fish sperm DNA, Bovine serum albumin were productsof Biophysics Institute of Chinese Academy of Science; Nitrocellulosemembrane (0.45 nm) was product of Amersham Co.

5. Method:

a). Duck Hepatitis B Virus Infection:

1-day-old Beijing duck was injected DHBV-DNA positive serum of Shanghaibrown-duck via vena cruralis with 0.2 ml/each duck. Blood was taken onthe seventh day after infected; serum was separated and stored at −70□to be detected.

b). Drug Treatment Experiment:

DHBV infected ducklings were randomly grouped (six ducklings/each group)to carry out drug treatment experiment after 7 days. The administrationgroups (W28D, W28F, W28M, C37) were set 1 dose group—50 mg/kgrespectively, and oral (intragastric) administered twice per day for 10days (Bid×10). The drugs were replaced with normal saline in viruscontrol group (DHBV). The positive drug was Lamivudine, which was oral(intragastric) administrated with 50 mg/kg, twice per day for 10 days.Blood was taken from the vena cruralis of the duck after 7 days later ofinfection, i.e. before drug administration (T0), the fifth day (T5) andthe tenth day after drug administration (T10), and the third day afterdrug withdrawal (P3), then serum was separated and stored at −70□ to bedetected.

c). Detection Method:

The above duck serum was taken and dotted on the membrane to measure thedynamic level of DHBV-DNA in the duck serum. According to theinstruction of nick translation kit, dot blot hybridization of duckserum and was carried out using ³² P labeled DHBV-DNA probe (purchasedfrom Promega Co.). And the dots were shown on the membrane byradioautography. The OD values (the filter was 490 nm) of the dots onthe membrane were measured by ELISA. DHBV-DNA density of the serum wascalculated and the OD values of the hybridized dot were used as thevalues of DHBV-DNA level.

6. Calculation of Drug Action:

a) The mean values (X±SD) of DHBV-DNA OD values of serum at differenttime of the ducks in each group were calculated, and the DNA levels ofserum at different time after drug administration (T5, T10) and thethird day after drug withdrawal (P3) for the ducks in each group andwere compared with the OD values before administration (T0) of the samegroup. The values of t1 and P1 were calculated by paired-t test. Thesignificance of the difference was analyzed, and the inhibitory effectof the drugs on the viral infection was estimated.

b). The inhibition ratio (%) to DHBV-DNA of serum at different timeafter administration (T5, T10) and the third day after drug withdrawal(P3) of the ducks in each group were calculated and illuminated in thefigure, and the dynamic state of the inhibition ratios of DHBV-DNA induck serum in each group were compared.

${{DNA}\mspace{14mu}{inhibition}\mspace{14mu}\%} = {\frac{\begin{matrix}{{{OD}\mspace{14mu}{value}\mspace{14mu}{before}\mspace{14mu}{administration}\mspace{14mu}\left( {T\; 0} \right)} -} \\{{OD}\mspace{14mu}{value}\mspace{14mu}{after}\mspace{14mu}{administration}\mspace{14mu}\left( {{T\; 5},{T\; 10},{P\; 3}} \right)}\end{matrix}}{{OD}\mspace{14mu}{Value}\mspace{14mu}{before}\mspace{14mu}{administration}\mspace{14mu}\left( {T\; 0} \right)} \times 100}$

c) The inhibitions % of DHBV-DNA at different time of the administrationgroup were compared with that at the same time of the virus controlgroup. The result was statistically treated with group t test, and thevalues of t2 and P2 were calculated. The significance of the differencewas analyzed, and the drug action was estimated.

7. Experimental Results:

FIG. 1 is the comparison of the inhibition ratio of DHBV-DNA level induck serum of the treating group and that of the control group withvirus infection, after the ducks infected by the duck hepatitis B viruswere orally (intragastric) administered.

With regard to 4 group with administrating drugs of the 50 mg/kg, afterthe ducks were orally (intragastric) administered twice per day for 10days, no change of the common condition and food intake of the duckswere observed.

In both paired and group analysis, the positive control drug,Lamivudine, of 50 mg/kg group showed inhibitory effect to DHBV-DNA,which means the experiment was tenable. 3 days after withdrawal of thedrugs, DHBV-DNA rebounded to the original level.

At the condition of the present experiment, for W28F 50 mg/kg, bothpaired analysis and group analysis of T5 and T10 showed inhibitoryeffect to DHBV-DNA with statistical significance. Though DHBV-DNA at P3rebounded, it did not reach the original level. For W28M 50 mg/kg group,though paired analysis of T10 and P3 showed inhibitory effect toDHBV-DNA with statistical significance, the group analysis did not showany effect with statistical significance. For W28D 50 mg/kg group, bothpaired analysis and group analysis T10 showed inhibitory effect toDHBV-DNA with statistical significance. For C37 50 mg/kg group, thepaired analysis and group analysis at each time point did not showinhibitory effect to DHBV-DNA.

The invention claimed is:
 1. A heterocyclic non-nucleoside compound represented by the following structure formula:

wherein each R₅ and R₆ independently is hydrogen atom, halogen atom, or phenyl; R₂ is linear or branched C₁-C₈ alkyl, C₃-C₈ cycloalkyl or benzyl; R₃ is one selected from the group consisting of C₁-C₆ hydroxyalkyl; C₁-C₄ alkyl substituted by C₁-C₄ alkoxycarbonyl; C₂-C₄ alkenyl substituted by C₁-C₄ alkoxycarbonyl;

 wherein each R′ and R″ independently is selected from the group consisting of halogenated C₁-C₁₀ alkyl, halogenated benzyl, benzyl substituted by C₁-C₁₀ alkyl, benzyl substituted by C₁-C₁₀ alkoxyl, benzyl substituted by C₁-C₁₀ alkylamino group, benzyl substituted by nitrile group, benzyl substituted by carboxyl, and benzyl substituted by C₁-C₁₀ alkyloxycarbonyl.
 2. The heterocyclic non-nucleoside compound according to claim 1, wherein said heterocyclic non-nucleoside compounds are one of the following compounds:


3. An antiviral pharmaceutical composition consisting of the heterocyclic non-nucleoside compound according to claim 1 as an active component and a pharmaceutically common adjuvant.
 4. A heterocyclic non-nucleoside compound selected from the following structure formula:


5. An antiviral pharmaceutical composition consisting of the heterocyclic non-nucleoside compound according to claim 4 as an active component and a pharmaceutically common adjuvant. 